TABLE 1.
Strains and plasmids
| Strain or plasmid | Relevant feature(s) | Reference or source |
|---|---|---|
| Strains | ||
| NM554 | F−araD139 Δ(ara-leu)7696 galE15 galK16 Δ(lac)X74 rpsL hsdR2 mcrA mcrB1 recA13 | 26 |
| BL21(DE3) | F−ompT lon hsdSBgal dcm(DE3) | 34 |
| 14:40-42 | F−araD139 Δlac(IPOZYA)U169 rpsL thi car-14(::Mu dlac Apr) pyrH42 | 28 |
| Plasmids | ||
| pET22b | ColE1 replication origin; T7 RNA polymerase-dependent expression vector; Apr | Novagen Inc. |
| pET24a | ColE1 replication origin; T7 RNA polymerase-dependent expression vector; Kmr | Novagen Inc. |
| pET24ma | p15A replication origin; T7 RNA polymerase-dependent expression vector; Kmr | 33a |
| pDIA17 | p15A replication origin; harbors the lacI gene; Cmr | 25 |
| pSL1 | Wild-type pyrH from E. coli in pET22b | This work |
| D168N | Mutated pyrH; D168N substitution | 8 |
| D174N | Mutated pyrH; D174N substitution | 8 |
| pSL2 | Truncated UMP kinase; fragment from residue 25 to 241 included | This work |
| pSL3 | Truncated UMP kinase; fragment from residue 50 to 241 included | This work |
| pHSP233 | Truncated UMP kinase; fragment from residue 113 to 241 included | This work |
| pSL4 | Truncated UMP kinase; fragment from residue 1 to 180 included | This work |
| pSL18 | Truncated UMP kinase; fragment from residue 1 to 160 included | This work |
| pSL19 | Truncated UMP kinase; fragment from residue 1 to 140 included | This work |
| pSL8 | Truncated UMP kinase; fragment from residue 1 to 120 included | This work |
| pSL11 | Wild-type UMP kinase from S. typhimurium SL5235 in pET24ma | This work |
| pSL13 | Wild-type UMP kinase from B. subtilis 168 in pET24a | This work |
| pHSP253 | Wild-type UMP kinase from H. influenzae 102-514T in pET24a | This work |